Evaluation of Matrix Issues in the Applicability of the Neuro-2a Cell Based Assay on the Detection of CTX in Fish Samples

Toxins (Basel). 2020 May 9;12(5):308. doi: 10.3390/toxins12050308.

Abstract

Ciguatoxins (CTXs) are a group of neurotoxins responsible for the syndrome ciguatera fish poisoning (CFP) as a result of the consumption of contaminated fish. The presence of these toxins has been detected around the Pacific, Caribbean and Indian coasts. Recent reports indicate the emergence of CFP in other geographic areas, in particular in European coasts, of the Canary Islands (Spain) and Madeira (Portugal). A neuroblastoma cell line of murine origin (N2a) has been applied to assay different groups of neurotoxins, acting on voltage-gated sodium channel (VGSC) of excitable cells, N2a-MTT. The great potential of N2a-MTT as a sensitive tool for the CTXs screening is clearly recognized, notably because it allows the detection of these toxins at levels below recommended as security levels. However, the complexity of the matrix is a critical point on the application of N2a-MTT, which needs to be evaluated. The aim of this work is to provide recommendations for an implemented N2a-MTT method for CTXs determination in fish that avoids matrix effects, particularly those related to high lipid content.

Keywords: ciguatoxins; matrix effect; neuroblastoma cell assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Assay*
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Ciguatoxins / analysis*
  • Ciguatoxins / pharmacology*
  • Fishes / metabolism*
  • Membrane Potentials
  • Mice
  • Neurons / drug effects*
  • Neurons / metabolism
  • Neurons / pathology
  • Ouabain / pharmacology
  • Veratridine / pharmacology
  • Voltage-Gated Sodium Channel Agonists / analysis*
  • Voltage-Gated Sodium Channel Agonists / pharmacology*
  • Voltage-Gated Sodium Channels / drug effects*
  • Voltage-Gated Sodium Channels / metabolism

Substances

  • Voltage-Gated Sodium Channel Agonists
  • Voltage-Gated Sodium Channels
  • Ciguatoxins
  • Ouabain
  • Veratridine